Name: a preclinical murine model of hepatic metastases
Uploaded: 2014-09-27T13:00:00.000+00:00
Duration: 6 min 51 s
Description: Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

KFとKOはKSと一緒に共同最初の著者である この作品は、ジョンズホプキンス医学部臨床科学者賞（LZ）、Viragh財団とジョンズ·ホプキンス大学をスキップViragh膵臓がんセンターは（EMJ、NIH K23 CA148964-01（LZ）、AHPBAフェローシップ（KS）によって部分的にサポートされていましたし、 LZ）、国立膵臓財団（LZ）、Lefkofskyファミリー財団（LZ）、胃腸癌のP50のCA062924（EMJ、LZ）、ラストガーテン財団（LZ）、およびソルゴールドマン膵臓がんセンター助成金のNCI SPORE（KS 、BHE、LZ）。

全ての動物実験は、ジョンズ·ホプキンス大学の動物実験委員会のガイドラインに適合し、動物を研究室のケアの評価と認定協会（AAALAC）のガイドラインに従って維持した。外科的処置は、毎時15空気交換の最小値を受け、手術室で、無菌条件下で行われる。すべての商品は、手順の間に、各マウスの間に70％エタノールで前の手順で、もう一度オートクレーブで滅菌されています。外科手術を生き残るために期待されていない全てのマウスは、麻酔下で、依然としてながら頸椎脱臼に続いて3分間、CO 2の下で安楽死させる。 1。細胞調製<ol><li> 2×10 7細胞/ mlで、リン酸緩衝生理食塩水で培養されたPanc02細胞を再懸濁し、そして氷上に保つ。 </li></ol> 2マウスの準備<ol><li>ケタミンの管理腹腔内分割用量で8〜12週齢のC57BL / 6雌マウスへの電子（100mg / kgの）キシラジン（10mg / kg）を混合した。 </li><li>つま先ピンチ引っ込め反射がマウスが完全に麻酔をかけていることを確実にするために廃止されていることを確認してください。 </li><li>完全にマウスを麻酔するために、必要に応じて（100mg / kgの）キシラジン（10mg / kgの）と混合ケタミンの追加の用量を投与する。 </li><li>マウスは麻酔下にある間、乾燥を防ぐために、マウスの眼にPuralube獣医軟膏を適用します。 </li><li>創傷の汚染を回避するために、マウスの手術領域を剃る。 </li><li>準備は、ヨウ素を70％エタノールで切開の肋骨下の領域を離れ、。 </li><li>無菌性を維持するために腹部に外科用ドレープを配置します。 </li></ol> 3開腹<ol><li>経皮、メスを用いて腹膜を介して左耳に沿って、左肋骨下の切開を加えます。 </li><li>同時デジタルPを適用することにより、切開部を通して脾臓エクスプレス切開の頭蓋および尾側の側面に沿って案内圧力スイッチ。 </li><li>脾臓の下端で脾臓の血管の位置を確認します。 </li><li>脾臓の極での脾臓の血管を損傷しないように注意しながら、脾臓の中心部に2ホライゾン中型結紮クリップを配置することによって、脾臓を割ります。 </li><li>汚染を避けるために戻って腹膜に脾臓の上極を配置します。 </li></ol> 4。腫瘍注射<ol><li> 26 Gのx 5/8 &quot;注射器の中にリン酸緩衝生理食塩水を150μlを描画します。 </li><li>注入時に常に直立位置に注射器を維持し、同じ注射器にPanc02100μlの（2×10 6）細胞を描画します。 </li><li> 70％エタノール溶液中に針を浸し。針は、無菌性を確保するために70％エタノールに浸漬する。 </li><li>注射器が常に直立させていることを確認されて露出した半脾臓にゆっくりと細胞を注入する。 syrinのベベルGEは脾臓で細胞を注入することを避けるために、脾臓カプセルを通じて常に観察されるべきである。脾臓や血管のホワイトニングは、注入時に観察されるべきである。 </li><li>脾臓を昇格し、脾臓の血管を閉塞するために、脾臓で1メディアホライゾンクリップを適用します。 </li><li>膵臓と脾臓血管の最も遠位の側面に小さなホライゾンクリップを適用します。 </li><li>ホライゾン·クリップの上にカットして半脾臓から膵臓と脾臓血管を連結する。 </li><li>その &#39;側にマウスを置き、蒸留水で切開を灌漑。 </li></ol>麻酔から5。腹部閉鎖およびリカバリ<ol><li> 4-0ランニングステッチで腹膜を閉じます。 </li><li>皮膚切開を閉じるために二から三の皮膚クリップを適用します。 </li><li>手術後の痛みを緩和するためには0.1mg / kgのブプレノルフィンまたは5〜10ミリグラム/ kgのカルプロフェンを皮下投与する。 </li><li> RECのための加熱パッド上でマウスを置きますオーヴェリーモバイルまで、とマウスは、通常の呼吸パターンを示しています。動物は、唯一の完全外科的手順から回復した後、他の動物の会社に戻ります。 </li></ol> 6。剖検審査及び肝臓の収穫<ol><li>手術後30〜60日の間、マウスは転移の臨床症状を示すように開始され、安楽死が必要になります。人道的な安楽死を必要とする転移性疾患の徴候は拡大された腹部および腹水の開発が含まれています。 CO 2の吸入によりマウスを安楽死させる。 </li><li>吸入後、頸椎脱臼を行います。 </li><li>動物は非反応性であることが決定された後、ハサミを使用して腹部を露出させるために腹膜を通って切開する。 </li><li>鉗子やハサミを使用して、静かに腹部から肝臓を切った。 </li><li> OCT化合物中の肝臓および液体窒素を用いたフラッシュ凍結を配置します。 -80℃で肝臓に保管してください。代わりに、ライブrは、埋め込まれた24時間およびパラフィンをRTで16％中性緩衝ホルマリンで固定することができる。 </li></ol>

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D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EMT+and+dissemination+precede+pancreatic+tumor+formation.">EMT and dissemination precede pancreatic tumor formation.</a> Cell. 148, 349-361 (2012).</li><li>Frese, K. K., Tuveson, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maximizing+mouse+cancer+models.">Maximizing mouse cancer models.</a> Nature reviews. Cancer. 7, 645-658 (2007).</li><li>Corbett, T. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effective+depletion+of+regulatory+T+cells+allows+the+recruitment+of+mesothelin-specific+CD8+T+cells+to+the+antitumor+immune+response+against+a+mesothelin-expressing+mouse+pancreatic+adenocarcinoma.">Effective depletion of regulatory T cells allows the recruitment of mesothelin-specific CD8 T cells to the antitumor immune response against a mesothelin-expressing mouse pancreatic adenocarcinoma.</a> Clinical and translational science. 1, 228-239 (2008).</li><li>Jain, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergistic+effect+of+a+granulocyte-macrophage+colony-stimulating+factor-transduced+tumor+vaccine+and+systemic+interleukin-2+in+the+treatment+of+murine+colorectal+cancer+hepatic+metastases.">Synergistic effect of a granulocyte-macrophage colony-stimulating factor-transduced tumor vaccine and systemic interleukin-2 in the treatment of murine colorectal cancer hepatic metastases.</a> Annals of surgical oncology. 10, 810-820 (2003).</li><li>Yoshimura, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+targeting+of+antitumor+immune+responses+with+engineered+live-attenuated+Listeria+monocytogenes.">Selective targeting of antitumor immune responses with engineered live-attenuated Listeria monocytogenes.</a> Cancer research. 66, 1096-1104 (2006).</li><li>Yoshimura, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+attenuated+Listeria+monocytogenes+effectively+treats+hepatic+colorectal+cancer+metastases+and+is+strongly+enhanced+by+depletion+of+regulatory+T+cells.">Live attenuated Listeria monocytogenes effectively treats hepatic colorectal cancer metastases and is strongly enhanced by depletion of regulatory T cells.</a> Cancer research. 67, 10058-10066 (2007).</li><li>Olino, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor-associated+antigen+expressing+Listeria+monocytogenes+induces+effective+primary+and+memory+T-cell+responses+against+hepatic+colorectal+cancer+metastases.">Tumor-associated antigen expressing Listeria monocytogenes induces effective primary and memory T-cell responses against hepatic colorectal cancer metastases.</a> Annals of surgical oncology. 19, 597-607 (2012).</li><li>Hingorani, S. 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著者らは、開示することは利害の関係する競合はありません。

半脾臓注入技術を経由して肝臓への膵臓癌転移のこの前臨床マウスモデルでは、肝臓への転移性疾患を有する患者の臨床的および免疫学的条件の多くを反映して説明した最初のモデルである。このモデルは、他のマウスモデルに比べていくつかの利点を提供しています。まず、膵臓癌のトランスジェニックモデルとは異なり、マウスのわずか30％が肝臓に転移を発症したおよび転移形成のタ?...

膵管腺癌（PDA）は、米国1における癌関連死の第四の主要な原因である。転移性PDAと患者のための5年生存率2-12％1です。膵臓癌のアジュバントおよびネオアジュバント化学療法はこれまで以上に、今日より効果的であるが、まだ新しい治療法のための絶望的な必要性が残っている。 新規治療薬の開発は、信頼性の前臨床動物モデルを必要とする。皮下腫瘍モデルを使用して簡単ですが、その欠点は、ヒトの癌で見られる腫瘍の進行および微小環境を転移し、模倣するこれらの腫瘍のできないことがあります。そのようなKrasのとp53ノックイン発癌性変異（KPCモデル）2とYFP系統トレーサー（PKCYモデル）とKPCモデルを含む遺伝子操作されたマウスなどの遺伝子操作されたモデルは、3 immunocompeteにおける自然発生腫瘍の研究を可能にntのマウス。加えて、腫瘍微小環境は変更され、より厳密にヒト腫瘍進行に見られるものに似ている。残念ながら、腫瘍発生のタイミングが困難実験的治療を研究することができ、これらのモデル内の変数である。さらに、これらのマウスの戻し交配は、常に実行可能ではない時間と財務コミットメントのかなりの量を必要とする。最後に、注入された異種移植のPDAマウスモデルは変更した免疫不全マウス、または存在しない腫瘍微小環境を必要とする。その結果、これらの前臨床モデルにおいて実証実験的治療の有効性は、多くの場合、ヒト臨床試験4において再現性がない。 このhemisplenectomyマウスモデルは、C57BL / 6マウス5、6から誘導される高度に腫瘍形成、メチルコラントレン誘発性膵臓腫瘍細胞株であるPanc02腫瘍細胞を利用する。さらに、他のネズミPDA細胞株は、KPCマイクから誘導eは、このモデルで使用することができる。 Schulick らは以前にhemisplenectomy技術を開発し、大腸癌転移7の同系マウスモデルを作成しました。このhemisplenectomyモデル（ 図1）は、新規の治療薬7-10の研究に自分自身を貸す免疫応答性マウスにおいて、一貫して同じ腫瘍接種を提供しています。

<table><tbody><tr><td>Cell culture media and components</td><td></td><td></td><td>will vary depending on cell line</td></tr><tr><td>Phosphate Buffered Saline</td><td>Gibco by Life Technologies</td><td>10010-023</td><td></td></tr><tr><td>15 ml centrifuge tubes</td><td>Corning</td><td>430052</td><td></td></tr><tr><td>Curved Operating Scissor</td><td>Roboz Surgical Store</td><td>RS-6835</td><td></td></tr><tr><td>Micro Dissecting Graefe Forceps</td><td>Roboz Surgical Store</td><td>RS-5130</td><td></td></tr><tr><td>Needle holder (regular)</td><td>World Precision Instruments, Inc</td><td></td><td></td></tr><tr><td>3-0 or 4-0 suture</td><td></td><td></td><td>courtesy of dept of surgery, Johns Hopkins</td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, small, angled</td><td>Teleflex</td><td>137085</td><td></td></tr><tr><td>Weck Horizon Open Ligating Clip Applier, medium, angled</td><td>Teleflex</td><td>237115</td><td></td></tr><tr><td>Weck Horizon medium ligating clips</td><td>Teleflex</td><td>002200</td><td></td></tr><tr><td>Weck Horizon small ligating clips</td><td>Teleflex</td><td>001200</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 3/8&quot; slip tip</td><td>Becton Dickinson</td><td>309625</td><td></td></tr><tr><td>Syringe, 1 cc, tb syringe, 5/8&quot; slip tip</td><td>Becton Dickinson</td><td>309597</td><td></td></tr><tr><td>9 mm Autoclip Applier</td><td>Mikron</td><td>427630</td><td></td></tr><tr><td>9 mm Autoclip</td><td>Becton Dickinson</td><td>427631</td><td></td></tr><tr><td>Heating pad</td><td>Sunbeam</td><td>CAT93</td><td></td></tr><tr><td>70% Ethanol</td><td></td><td></td><td>sold by numerous vendors</td></tr><tr><td>Ketamine Hydrochloride&amp;nbsp;</td><td>Hospira</td><td>22395DD</td><td></td></tr><tr><td>Xylazine hydrochloride</td><td>Sigma</td><td>X1251</td><td></td></tr></tbody></table>

a preclinical murine model of hepatic metastases

多数のマウスモデルは、ヒト癌を研究し、癌治療及び開発の理解を進めるために開発されてきた。ここで、同系マウスの膵臓腫瘍細胞の半脾臓注射を介して、肝転移の前臨床マウス膵臓腫瘍モデルについて説明する。このモデルは、肝臓への転移性疾患を有する患者における臨床症状の多くを模倣。マウスは、一貫して転移過程、実験的治療試験、および腫瘍免疫学研究の調査を可能に肝臓への転移を開発しています。

半脾臓に注入Panc02腫瘍細胞技術が正しく行われた場合、肺および腹膜転移が観察されない一方（ 図1）マウスの100％においてフォームの肝臓転移、。この技術は、複数の独立した実験においてPanc02細胞を使用して100以上のマウスに対して行われており、再現性、すべての未処置のマウスはhemisplenectomy手順（ 図2）以下の30と60日の間の肝転移で死亡する。未処理および?...

Watch this Scientific Journal Video about A Preclinical Murine Model of Hepatic Metastases at JoVE.com

A Preclinical Murine Model of Hepatic Metastases

肝転移の前臨床マウスモデルは、半脾臓注入技術を介して行う。

肝転移の前臨床マウスモデル

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

a-preclinical-murine-model-of-hepatic-metastases

Research

JoVE Journal

Medicine

28.5K ビュー.  The Johns Hopkins University School of Medicine. 肝転移の前臨床マウスモデルは、半脾臓注入技術を介して行う。 

ビデオ: A Preclinical Murine Model of Hepatic Metastases

Murine Bioluminescent Hepatic Tumour Model

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are administered specifically to the liver to induce a localised liver tumour, via mobilisation of the spleen and splitting into two, leaving intact the vascular pedicle for each half of the spleen. Lewis lung carcinoma cells that constitutively express the firefly luciferase gene (luc1) are inoculated into one hemi-spleen which is then resected 10 minutes later. The other hemi-spleen is left intact and returned to the abdomen. Liver tumour growth can be monitored by bioluminescence imaging using the IVIS whole body imaging system. Quantitative imaging of tumour growth using IVIS provides precise quantitation of viable tumour cells. Tumour cell death and necrosis due to drug treatment is indicated early by a reduction in the bioluminescent signal. This mouse model allows for investigating the mechanisms underlying metastatic tumour-cell survival and growth and can be used for the evaluation of therapeutics of liver metastasis. 

This article describes a procedure for the induction of orthotopic bioluminescent liver tumours in mice, and subsequent analysis of tumour growth confined to the liver using live whole body luminescence imaging.

マウス生物発光肝腫瘤モデル

This video describes the establishment of liver metastases in a mouse model that can be subsequently analysed by bioluminescent imaging. Tumour cells are ...

医学

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. However, little is known about the heterogeneity of liver metastases, and animal models capable of evaluating the development of individual metastatic colonies are needed. Here, we present an advanced model of hepatic metastases that provides the ability to quantitatively visualize the development of individual tumor clones in the liver and estimate their growth kinetics and colonization efficiency. We generated a panel of monoclonal derivatives of HCT116 human colorectal cancer cells stably labeled with luciferase and tdTomato and possessing different growth properties. With a splenic injection followed by a splenectomy, the majority of these clones are able to generate hepatic metastases, but with different frequencies of colonization and varying growth rates. Using the In Vivo&#160;Imaging System (IVIS), it is possible to visualize and quantify metastasis development with in vivo luminescent and ex vivo fluorescent imaging. In addition, Diffuse Luminescent Imaging Tomography (DLIT) provides a 3D distribution of liver metastases in vivo. Ex vivo fluorescent imaging of harvested livers provides quantitative measurements of individual hepatic metastatic colonies, allowing for the evaluation of the frequency of liver colonization and the growth kinetics of metastases. Since the model is similar to clinically observed liver metastases, it can serve as a modality for detecting genes associated with liver metastasis and for testing potential ablative or adjuvant treatments for liver metastatic disease.

The ability of metastatic clones to colonize distant sites depends on their proliferation capacity and/or their ability to survive in the host microenvironment without significant proliferation. Here, we present an animal model that allows quantitative visualization of both types of liver colonization by metastatic clones.

肝臓における大腸転移の高度な動物モデル：イメージング技術および転移性クローンのプロパティ

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. ...

がん研究

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study end-points. Thus, studies focused on elucidating mechanisms of late-stage de novo metastasis are compromised, as are studies examining efficacy of anti-cancer therapies targeting mediators of metastasis in the adjuvant setting. Numerous murine mammary cancer models have been developed via targeted expression of dominant oncoproteins to mammary epithelial cells yielding models variably mimicking histopathologic and transcriptome-defined breast cancer subtypes common in women1. While much has been learned regarding the biology of mammary carcinogenesis with these models, their utility in identifying molecules regulating growth of late-stage metastasis are compromised as mice are typically euthanized at earlier time points due to significant primary tumor burden. Moreover, since a significant percentage of women diagnosed with breast cancer receive adjuvant therapy after surgical resection of primary tumors and prior to presence of detectable metastatic disease, preclinical models of de novo metastasis are urgently needed as platforms to evaluate new therapies aimed at targeting metastatic foci. To address these deficiencies, we developed a murine model of de novo mammary cancer metastasis, wherein primary mammary tumors are surgically resected, and metastatic foci subsequently develop over a 115 day post-surgical period. This long latency provides a tractable model to identify functionally significant regulators of metastatic progression in mice lacking primary tumor, as well as a model to evaluate preclinical therapeutic efficacy of agents aimed at blocking functionally significant molecules aiding metastatic tumor survival and growth.

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

Pre-clinical models evaluating adjuvant therapy targeting breast cancer metastasis are lacking. To address this, we developed a murine model of de novo pulmonary mammary adenocarcinoma metastasis, wherein therapies administered in the adjuvant setting (post surgical resection of primary tumors) can be evaluated for efficacy in impacting previously seeded pulmonary metastases.

前臨床マウスモデルのための外科手術手順および方法論<em&gt;デ・ノヴォ</em&gt;乳がん転移

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study ...

A Portal Vein Injection Model to Study Liver Metastasis of Breast Cancer

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast cancer metastasis, and results in poor outcomes with median survival rates of only 4.8 - 15 months. Current rodent models of breast cancer metastasis, including primary tumor cell xenograft and spontaneous tumor models, rarely metastasize to the liver. Intracardiac and intrasplenic injection models do result in liver metastases, however these models can be confounded by concomitant secondary-site metastasis, or by compromised immunity due to removal of the spleen to avoid tumor growth at the injection site. To address the need for improved liver metastasis models, a murine portal vein injection method that delivers tumor cells firstly and directly to the liver was developed. This model delivers tumor cells to the liver without complications of concurrent metastases in other organs or removal of the spleen. The optimized portal vein protocol employs small injection volumes of 5 - 10 &#956;l, &#8805; 32 gauge needles, and hemostatic gauze at the injection site to control for blood loss. The portal vein injection approach in Balb/c female mice using three syngeneic mammary tumor lines of varying metastatic potential was tested; high-metastatic 4T1 cells, moderate-metastatic D2A1 cells, and low-metastatic D2.OR cells. Concentrations of &#8804; 10,000 cells/injection results in a latency of ~ 20 - 40 days for development of liver metastases with the higher metastatic 4T1 and D2A1 lines, and &#62; 55 days for the less aggressive D2.OR line. This model represents an important tool to study breast cancer metastasis to the liver, and may be applicable to other cancers that frequently metastasize to the liver including colorectal and pancreatic adenocarcinomas.

A surgical procedure was developed to deliver mammary tumor cells to the murine liver via portal vein injection. This model permits investigation of late stages of liver metastasis in a fully immune competent host, including tumor cell extravasation, seeding, survival, and metastatic outgrowth in the liver.

乳癌の肝転移を研究するためのポータル静脈注射モデル

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast ...

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded elucidation of the underlying mechanisms and development of innovative immunotherapeutic strategies. To develop an ideal animal model recapitulating human HCC, immunocompetent male C57BL/6J mice first receive a carbon tetrachloride (CCl4) injection to induce liver fibrosis, then receive histologically-normal oncogenic hepatocytes from young male SV40 T antigen (TAg)-transgenic mice (MTD2) by intra-splenic (ISPL) inoculation. Androgen generated in recipient male mice at puberty initiates TAg expression under control of a liver-specific promoter. As a result, the transferred hepatocytes become cancer cells and form tumor masses in the setting of liver fibrosis/cirrhosis. This novel model mimics human HCC initiation and progression in the context of liver fibrosis/cirrhosis and reflects the most typical features of human HCC including immune dysfunction.

Here, we describe the development of a clinically relevant murine model of liver cancer recapitulating the typical immune features of hepatocellular cancer (HCC).

肝炎症と線維症の設定に起因する肝細胞癌の腫瘍性肝細胞誘発性オルコトピックマウスモデル

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...

Murine Model of Metastatic Liver Tumors in the Setting of Ischemia Reperfusion Injury

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce multiple tissue and organ damage. Despite recent advances and therapeutic approaches, the overall morbidity has remained unsatisfactory especially in patients with underlying parenchymal abnormalities. In the context of aggressive cancer growth and metastasis, surgical I/R is suspected to be the promoter regulating tumor recurrence. This article aims to describe a clinically relevant murine model of liver I/R and colorectal liver metastasis. In doing so, we aim to assist other investigators in establishing and perfecting this model for their routine research practice to better understand the effects of liver I/R on promoting liver metastases.

We describe in detail a clinically relevant colorectal cancer liver metastases (CRLM) tumor model and the influence of liver ischemia reperfusion (I/R) in tumor growth and metastasis. This model can help to better understand the mechanisms underlying surgery-induced promotion of liver metastatic growth.

虚血多灌流損傷の設定における転移性肝腫瘍のマウスモデル

Liver ischemia and reperfusion (I/R) injury, a common clinical challenge, remains an inevitable pathophysiological process that has been shown to induce ...

Portal Vein Injection of Colorectal Cancer Organoids to Study the Liver Metastasis Stroma

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the tumor microenvironment, play a crucial role in metastatic CRC progression and predict poor patient prognosis. However, there is a lack of satisfactory mouse models to study the crosstalk between metastatic cancer cells and CAFs. Here, we present a method to investigate how liver metastasis progression is regulated by the metastatic niche and possibly could be restrained by stroma-directed therapy. Portal vein injection of CRC organoids generated a desmoplastic reaction, which faithfully recapitulated the fibroblast-rich histology of human CRC liver metastases. This model was tissue-specific with a higher tumor burden in the liver when compared to an intra-splenic injection model, simplifying mouse survival analyses. By injecting luciferase-expressing tumor organoids, tumor growth kinetics could be monitored by in vivo imaging. Moreover, this preclinical model provides a useful platform to assess the efficacy of therapeutics targeting the tumor mesenchyme. We describe methods to examine whether adeno-associated virus-mediated delivery of a tumor-inhibiting stromal gene to hepatocytes could remodel the tumor microenvironment and improve mouse survival. This approach enables the development and assessment of novel therapeutic strategies to inhibit hepatic metastasis of CRC.

Portal vein injection of colorectal cancer (CRC) organoids generates stroma-rich liver metastasis. This mouse model of CRC hepatic metastasis represents a useful tool to study tumor-stroma interactions and develop novel stroma-directed therapeutics such as adeno-associated virus-mediated gene therapies.

肝転移間質を研究するための結腸直腸癌オルガノイドの門脈注入

Hepatic metastasis of colorectal cancer (CRC) is a leading cause of cancer-related death. Cancer-associated fibroblasts (CAFs), a major component of the ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

Our laboratory has developed a novel orthotopic implantation model of human prostate cancer (PCa). As PCa death is not due to the primary tumor, but rather the formation of distinct metastasis, the ability to effectively model this progression pre-clinically is of high value. In this model, cells are directly implanted into the ventral lobe of the prostate in Balb/c athymic mice, and allowed to progress for 4-6 weeks. At experiment termination, several distinct endpoints can be measured, such as size and molecular characterization of the primary tumor, the presence and quantification of circulating tumor cells in the blood and bone marrow, and formation of metastasis to the lung. In addition to a variety of endpoints, this model provides a picture of a cells ability to invade and escape the primary organ, enter and survive in the circulatory system, and implant and grow in a secondary site. This model has been used effectively to measure metastatic response to both changes in protein expression as well as to response to small molecule therapeutics, in a short turnaround time.

This orthotopic model of human prostate cancer allows for quantification of tumor size, circulating tumor cells, and formation of distinct metastasis to the lung. As cells must escape the primary organ, enter the blood stream, and implant into a secondary site, this model effectively recapitulates the scenario in humans.

ヒト前立腺癌転移の同所マウスモデル

Our laboratory has developed a novel orthotopic implantation model of  human prostate cancer (PCa). As PCa death is not due to the primary tumor, but  ...

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

肝細胞癌の同所性モデルマウスにおける非侵襲的PET/MRイメージング

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of cancer metastasis are still unclear. Animal models are essential for elucidating the mechanisms and for evaluating novel strategies for the treatment of metastatic cancers. Here, an in-depth description of a &#8220;patient-like&#8221; orthotopic syngeneic mouse model for exploring the mechanisms of metastasis of solid organ tumors is provided. The survival surgical implantation of BNL 1ME A.7R.1 mouse hepatocellular carcinoma cells directly into the liver (the organ of origin) of the inbred wild-type immune competent laboratory mouse strain, BALB/c is described. The success and reproducibility of this methodology recommends it for widespread use in elucidating the biological mechanisms of solid organ cancer metastasis.

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a &#8220;patient-like&#8221; orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

肝細胞癌転移の「患者様」同所同系マウスモデル

The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...